Different methods are well established and widespread used for protein purification, such as affinity chromatography with microbial proteins (e.g. protein A or protein G affinity chromatography), ion exchange chromatography (e.g. cation exchange (carboxymethyl resins), anion exchange (amino ethyl resins) and mixed-mode exchange), thiophilic adsorption (e.g. with beta-mercaptoethanol and other SH ligands), hydrophobic interaction or aromatic adsorption chromatography (e.g. with phenyl-sepharose, aza-arenophilic resins, or m-aminophenylboronic acid), metal chelate affinity chromatography (e.g. with Ni(II)- and Cu(II)-affinity material), size exclusion chromatography, and electrophoretical methods (such as gel electrophoresis, capillary electrophoresis) (Vijayalakshmi, M. A., Appl. Biochem. Biotech. 75 (1998) 93-102).
The industrial purification of pharmaceutical antibodies, especially the development, operation and validation of chromatography processes is reported by Fahrner, R. L., et al., in Biotechnol. Gen. Eng. Rev. 18 (2001) 301-327. Follman, D. K. and Fahrner, R. L. (J. Chrom. A 1024 (2004) 79-85) report a factorial screening of antibody purification processes using three chromatography steps without protein A. The capture of human monoclonal antibodies from cell culture supernatant by ion exchange media exhibiting high charge density is reported by Necina, R., et al. (Biotechnol. and Bioeng. 60 (1998) 689-698). Protein purification by ion exchange chromatography is reported in WO 99/057134. In WO 2004/076485 antibody purification by protein A and ion exchange chromatography is reported. In U.S. Pat. No. 5,429,746 antibody purification is reported. Protein purification is reported in WO 2003/066662.
WO 2006/125599 reports a method for the purification of antibodies. Antibody purification by protein A and ion exchange chromatography is reported in WO 2004/076485.